Pregnant Female Mice Research Articles

Chronic exposure of bisphenol-A impairs cognitive function and disrupts hippocampal insulin signaling pathway in male mice

Bisphenol-A (BPA), a well-known estrogenic endocrine disruptor, is generally applied to turn out plastic consumer products. Available data have manifested that exposure to BPA can trigger insulin resistance. Hence, the purpose of the actual study was to consider the impacts of BPA exposure on cognitive function and insulin signaling pathway in the hippocampus of male offspring mice. For this purpose, the pregnant female mice were treated either vehicle (0.1% ethanol) or BPA (0.01, 0.1, and 1 µg/mL) via drinking water from day 1 of gestation until delactation (D1-PND21, newborn exposure). Afterward, the three-week-old male offspring mice took orally with the same doses of BPA for nine weeks (PND84). The behavioral tests, blood sugar level, histological observation, transcriptome sequencing, glucose transporter 4 (GLUT4), and hippocampal insulin signaling pathway were checked for the male offspring mice at 13 weeks of age (PND91). Our data indicated that BPA exposure impaired cognitive function, disrupted the hippocampal regular cell arrangement, increased blood glucose levels, disturbed the insulin signaling pathway including phosphorylated insulin receptor substrate1 (p-IRS1), protein kinase B (p-AKT), and glycogen synthase kinase 3β (p-GSK3β). At the same time, the mRNA and protein expressions of GLUT4 were markedly down-regulated in the BPA-exposed groups. To sum up, it has been suggested from these results that BPA has detrimental effects on the insulin signaling pathway, which might subsequently be conducive to the impairment of cognitive function in the adult male offspring mice. Therefore, BPA exposure might in part be an element of risk for the long-term neurodegeneration in male offspring mice.

Lack of Vesicular Zinc Does Not Affect the Behavioral Phenotype of Polyinosinic:Polycytidylic Acid-Induced Maternal Immune Activation Mice.

Zinc is important in neural and synaptic development and neuronal transmission. Within the brain, zinc transporter 3 (ZnT3) is essential for zinc uptake into vesicles. Loss of vesicular zinc has been shown to produce neurodevelopmental disorder (NDD)-like behavior, such as decreased social interaction and increased anxiety- and repetitive-like behavior. Maternal immune activation (MIA) has been identified as an environmental factor for NDDs, such as autism spectrum disorders (ASDs) and schizophrenia (SZ), in offspring, which occurs during pregnancy when the mother’s immune system reacts to the exposure to viruses or infectious diseases. In this study, we investigated the interaction effect of a genetic factor [ZnT3 knockout (KO) mice] and an environmental factor (MIA). We induced MIA in pregnant female (dams) mice during mid-gestation, using polyinosinic:polycytidylic acid (polyI:C), which mimics a viral infection. Male and female ZnT3 KO and wild-type (WT) offspring were tested in five behavioral paradigms: Ultrasonic Vocalizations (USVs) at postnatal day 9 (P9), Open Field Test, Marble Burying Test, three-Chamber Social Test, and Pre-pulse Inhibition (PPI) in adulthood (P60–75). Our results indicate that loss of vesicular zinc does not result in enhanced ASD- and SZ-like phenotype compared to WT, nor does it show a more pronounced phenotype in male ZnT3 KO compared to female ZnT3 KO. Finally, MIA offspring demonstrated an ASD- and SZ-like phenotype only in specific behavioral tests: increased calls emitted in USVs and fewer marbles buried. Our results suggest that there is no interaction between the loss of vesicular zinc and MIA induction in the susceptibility to developing an ASD- and SZ-like phenotype.

Maternal iron supplementation during pregnancy affects placental function and iron status in offspring

BackgroundIron deficiency and overload during pregnancy damage to maternal and fetal health. Placenta as an organ for the transport of nutrients between mother and fetus protects fetus from the harmful effects of iron deficiency and iron overload through regulation of placental iron homeostasis. MethodsTo determine the effect of dietary iron supplementation during pregnancy on reproduction and the mechanism of placental iron regulation, we designed dietary high iron (HI: 344 mg/kg), medium iron (MI: 40 mg/kg), low iron (LI: 2 mg/kg) groups of pregnant female mice fed ferrous citrate 2 weeks before mating to 18.5 days of gestation. ResultsWe find dietary iron supplementation during pregnancy effect maternal liver iron, placental iron, hemoglobin and fetal iron. Dietary iron significantly improves reproductive performance as litter weight and fetal weight. Correlation analysis suggest placental iron increased with liver iron, higher and lower liver iron is not conducive to the accumulation of fetal iron, placental iron deficiency and excess reduce litter weight. Placental transcriptome analysis revealed DEGs with the same trend in HI and LI groups compared with MI group, dietary iron may change biology process of ion transport and gland development in placenta. Granzyme may affect the placental trophoblast structure prior to delivery with iron overload uniquely. ConclusionThis research highlights the importance of moderate iron supplements in pregnancy due to damage of reproduction by affecting placental function under different dose of maternal iron supplementation.

Congenital anomalies and spontaneous abortion in mice resulting from the use of escitalopram.

Escitalopram (ESC) use during pregnancy has not been associated with teratogenic effects in fetuses. To investigate whether ESC administered during pregnancy in mice induces maternal toxicity and teratogenicity in offspring. Treated mice groups G1 and control G0 (n =15 per group). Administration of ESC (G1) and saline solution (G0) during pregnancy and euthanasia on the 18thday. Pregnant female mice were treated with ESC (20mg/kg, via gavage) or saline solution (control group) from the 5th to the 17thday of gestation, when implantation was consolidated. During intraembryonic development until the day before delivery, the drug had an influence on the development of alterations from its maintenance in the uterine environment and its development to the disturbance causing skeletal or visceral malformations. The intrauterine development parameters that were altered by ESC treatment were: number of resorptions (G0: [0.93±0.24]); G1: [3.33±0.51]), post-implantation loss (G0: [3.95±1.34], G1: [13.75±3.62]) and reduced fetal viability: [97.30±1.00]; G1: [81.09±6.22]). Regarding fetal formation, the treated group had visceral malformations with a significant frequency: cleft palate (G0: [1.0%], G1: [11.86%]) and reduced kidneys (G0: [0%]; G1: [10.17%]). Regarding skeletal malformations, a higher frequency was observed in the following parameters: incomplete supraoccipital ossification (G0: [0%], G1: [15.25]), absence of ribs (G0: [0%], G1 (G0: [0%], G1 [15.25%]) and absence of one or more of the foot phalanges (G0: [1.0%]; 64%]). Results indicate that ESC is an embryotoxic and teratogenic drug. Until further studies are performed, greater caution is necessary in prescribing the drug to pregnant women.

Hyperprolactinemia Modifies Extracellular Matrix Components that are Associated with Fibrillogenesis of Collagen Fibers in Harderian Glands of the Non- And Pregnant Female Mice

Hyperprolactinemia Modifies Extracellular Matrix Components that are Associated with Fibrillogenesis of Collagen Fibers in Harderian Glands of the Non- And Pregnant Female Mice

Effect of Corn Silk Aqueous Extract on Brown Adipose Tissue of Embryos and Neonates of Diabetic Pregnant Mice: A Histological Study.

Many congenital malformations are seen increasingly, due to diabetic mothers causing a burden on health systems. Corn silk (CS) extract has been used as a natural hypoglycemic treatment. However, its teratogenic safety was not studied. Therefore, in this study, we examine the effect of CS aqueous extract on fetuses, offspring of normal and diabetic female mice treated with CS aqueous extract. Pregnant female mice were divided into two groups diabetic and nondiabetic. Then, each of these groups was divided into control and treated. A daily dose of 4 g/kg of CS aqueous extract was given orally to the treated groups, control groups were given distilled water. The collection of samples was at day 16.5 of pregnancy, and neonates. Brown adipose tissue (BAT) in the sections of the preserved sample was examined. BAT areas were measured from 10 samples of each treatment age group in 2 sections. Data were analyzed with one-way ANOVA, then, two-independent sample test (Mann-Whitney) was done to test the significance of differences between groups. The BAT areas were negatively affected by diabetes and the extract. Both the extract and diabetes caused an increase in fat accumulation in the adipocytes with varying degrees. This study showed for the first time to our knowledge that the use of CS aqueous extract during pregnancy affected BAT organization and area, and that the used dose did not decrease the malformations caused by diabetes. More studies with different doses should be investigated.

Prenatal exposure to α-pinine improves reflexive motor behaviours in mice offspring.

α-pinene is a well-known compound representative of the monoterpenes group with a wide range of pharmacological activities. This article aims to determine effects of the prenatal exposure to α-pinene on reflexive motor behaviours in mice offspring. Forty pregnant female NMRI mice (8-10weeks old) were allocated into four groups. Group 1 served as control and groups 2-4 were intraperitoneally (i.p.) injected α-pinene (0.1, 0.5 and 1mg/kg) on 5, 8, 11, 14 and 17 days of gestation (GD). The control group was injected with saline at the same days. Following delivery, 20 pups from each litter were selected and reflexive motor behaviours determined using ambulation, hindlimb foot angle, surface righting, hindlimb strength, grip strength, front-limb suspension and negative geotaxis tests. Based on the findings of the present study, maternal exposure to α-pinene increased ambulation score, hind-limb suspension score, grip strength, front-limb suspension compared with the control group (P<0.05). Also, prenatal exposure to α-pinene decreased surface righting, hind-limb foot angle and negative geotaxis in mice offspring compared with the control group (P<0.05). α-pinene (0.1, 0.5 and 1mg/kg) decreased blood MDA and increased SOD and GPx levels in mice offspring (P<0.05). These results suggested α-pinene exposure during pregnancy has positive effect on reflexive motor behaviours in mice offspring possibly due to its antioxidant properties.

Ghrelin and Cannabinoid Functional Interactions Mediated by Ghrelin/CB1 Receptor Heteromers That Are Upregulated in the Striatum From Offspring of Mice Under a High-Fat Diet.

There is evidence of ghrelinergic-cannabinoidergic interactions in the central nervous system (CNS) that may impact on the plasticity of reward circuits. The aim of this article was to look for molecular and/or functional interactions between cannabinoid CB1 and ghrelin GHS-R1a receptors. In a heterologous system and using the bioluminescence resonance energy transfer technique we show that human versions of cannabinoid CB1 and ghrelin GHS-R1a receptors may form macromolecular complexes. Such receptor heteromers have particular properties in terms of CB1/Gi-mediated signaling and in terms of GHS-R1a-Gq-mediated signaling. On the one hand, just co-expression of CB1R and GHS-R1a led to impairment of cannabinoid signaling. On the other hand, cannabinoids led to an increase in ghrelin-derived calcium mobilization that was stronger at low concentrations of the CB1 receptor agonist, arachidonyl-2’-chloroethylamide (ACEA). The expression of CB1-GHS-R1a receptor complexes in striatal neurons was confirmed by in situ proximity ligation imaging assays. Upregulation of CB1-GHS-R1a- receptor complexes was found in striatal neurons from siblings of pregnant female mice on a high-fat diet. Surprisingly, the expression was upregulated after treatment of neurons with ghrelin (200 nM) or with ACEA (100 nM). These results help to better understand the complexities underlying the functional interactions of neuromodulators in the reward areas of the brain.

THE EFFECT OF PREGNANCY MILK ON THE EXPRESSION OF KALLIKREIN RELATED PEPTIDASE-4 (KLK-4) AND COLLAGEN TYPE 1 (Coll-1) IN AMELOGENESIS

Background: Tooth development during embryonic period is a complex process and requires adequate nutrients for the formation of healthy dental tissues. Kallikrein-related peptidase-4 (KLK-4) and collagen type 1 (Coll-1) are serine proteinases secreted by ameloblast during the transition and maturation stages of the amelogenesis processes, functioning to degrade the protein matrixes, so that the enamel can reach its final hardness. Pregnancy milk contains various nutrients expected to increase the KLK-4 expression of ameloblast cells in tooth development processes Purpose: This study aimed at determining the influence of pregnancy milk on the KLK-4 and collagen type 1 (Coll-1) expression of ameloblast cells in the tooth development processes.study Method The research subjects comprised of 10 pregnant female mice (Mus Musculus L.) that were divided into: control group (given sterile aquadest) and treatment group (given pregnancy milk + sterile aquadest) for 18 days followed by the collection of the tooth germ. The specimens were then stained using Imunnohistochemistry to see the KLK-4 and Coll-1 expressions. The data were analyzed using a pathway analysis. Result: The average KLK-4 and Coll-1 expression in the treatment group were higher than those in the control group. Based the pathway analysis, there were direct correlation of Pregnancy milk with Coll-1 expression and that with KLK-4 and Coll-1 expression as well as indirect correlation of pregnancy milk with KLK-4 expression. Conclusion: Pregnancy milk influences the Kallikrein-related peptidase-4 (KLK-4) and Coll-1 expression of ameloblast cells in the tooth development of the mice’s fetusesKeywords: Coll-1 pregnancy milk, Kallikrein-related peptidase-4 (KLK-4), Tooth development

Sex Differences in Embryonic Gonad Transcriptomes and Benzo[a]pyrene Metabolite Levels After Transplacental Exposure.

Polycyclic aromatic hydrocarbons like benzo[a]pyrene (BaP) are generated during incomplete combustion of organic materials. Prior research has demonstrated that BaP is a prenatal ovarian toxicant and carcinogen. However, the metabolic pathways active in the embryo and its developing gonads and the mechanisms by which prenatal exposure to BaP predisposes to ovarian tumors later in life remain to be fully elucidated. To address these data gaps, we orally dosed pregnant female mice with BaP from embryonic day (E) 6.5 to E11.5 (0, 0.2, or 2 mg/kg/day) for metabolite measurement or E9.5 to E11.5 (0 or 3.33 mg/kg/day) for embryonic gonad RNA sequencing. Embryos were harvested at E13.5 for both experiments. The sum of BaP metabolite concentrations increased significantly with dose in the embryos and placentas, and concentrations were significantly higher in female than male embryos and in embryos than placentas. RNA sequencing revealed that enzymes involved in metabolic activation of BaP are expressed at moderate to high levels in embryonic gonads and that greater transcriptomic changes occurred in the ovaries in response to BaP than in the testes. We identified 490 differentially expressed genes (DEGs) with false discovery rate P-values < 0.05 when comparing BaP-exposed to control ovaries but no statistically significant DEGs between BaP-exposed and control testes. Genes related to monocyte/macrophage recruitment and activity, prolactin family genes, and several keratin genes were among the most upregulated genes in the BaP-exposed ovaries. Results show that developing ovaries are more sensitive than testes to prenatal BaP exposure, which may be related to higher concentrations of BaP metabolites in female embryos.

Morphological Study of the Effects of Caffeine Beverages Can Cause Birth Defects on Swiss White Mice Embryos

The current study was conducted to identify the effects of different doses of Arabic coffee, black tea and Coca-Cola syrup on the possibility of birth defects in Swiss white mice embryos. (25) pregnant female mice were used in the study, divided into five experimental groups, including the control group dosed with distilled water, and four experimental groups were dosed with concentrations (4000-2500-2000-1000) mg/kg body weight for Arabic coffee and black tea and (2,3,4,6 ml/kg) for Coca-Cola syrup; once a day, from the seventh day until the eighteenth day of pregnancy. The results of the current study showed cases of abortion and fetal death at varying degrees and many abnormalities in white mouse embryos due to the different concentrations of the beverages used were characterized by the appearance of elongated and enlarged embryos. The elongation and deformation of the fetus, And the embryos mutated. Small embryos folded like a ball and curved in the shape of letter C, and the severity of the deformities increased by the appearance of mutated embryos resembling the body of a fish and deformed embryos similar to a water mermaid and not fully developed and the appearance of the head in the shape of a blurred triangle, as well as forward with a pointed end resembling a bird's beak, and brain deformation through depression in the posterior cranial region of the skull, and for the first time a new deformation is a cerebral Meningomyelocele in the form of a cystic tumor in the posterior brain region.

Lead acetate induced toxicities and antitoxic effect of Vitamin E and selenium in mice

Background: The experiment was conducted to evaluate the effect of different dosages of lead in the hematological parameters of mice and to observe the antitoxic role of Vitamin E and Selenium in induced lead toxicities. Moreover, the toxic effect of lead in the reproduction of female mice was also examined. Methods: A total of 72 (48 male and 24 female) Swiss albino mice were used in the experiment. After adaptation, 42 male mice were selected for hematological studies and divided into seven groups (n=6) where Group A represented healthy control mice and Group B, C, and D were treated with lead acetate at the rate of 0.5mg/kg, 1mg/kg and 2mg/kg respectively. Similarly, three other groups B+, C+, and D+ were treated with lead acetate plus Vitamin E and Selenium at the rate of 2ml per liter drinking water. A total of 24 female mice were divided into four groups (n=6), group E represented control mice and Group F, G, and H were treated with lead acetate at the rate of 0.5mg/kg, 1mg/kg, and 2mg/kg for three weeks followed by matting and treatment was continued for another one week of gestation. Blood sample was analyzed from the hematological study group. Result: The lead treatment caused a dose-dependent decrease in the value of Hb and PCV significantly whereas the value of TEC and TLC were significantly decreased in Group C and D in relation to Control. The value of ESR increased significantly in a dose-dependent manner in Group D in relation to Control whereas MCV and MCH values were significantly decreased than that of control. The value of TEC, Hb, PCV, ESR, and TLC improved in the Lead plus Vitamin E- Selenium treated group as compared to the Lead treated group but, only Group C+ showed significant improvement as compared to Group C. The value of neutrophil and monocyte were significantly decreased were as lymphocyte and eosinophil were significantly increased relative to control. There was a dose-dependent effect of lead in pregnancy of female mice with the highest effect (premature delivery and infant mortality) on high dose treated mice. Conclusion: It can be concluded that lead has a great impact on hematological parameters and has an effect on various systems of the body. Premature birth and abortion are major effects of lead toxicity. Our results suggest that hemolysis of RBC and or impairment of erythropoiesis may be caused by lead toxicity and the hematological values can be restored by the use of Vitamin E plus Selenium.

Influence of leptin administration to pregnant female mice on obesity development, taste preferences, and gene expression in the liver and muscles of their male and female offspring.

. The consumption of food rich in sugar and fat provokes obesity. Prenatal conditions have an impact on taste preferences and metabolism in the adult offspring, and this impact may manifest differently in different sexes. An increase in blood leptin level in pregnant females reduces the risk of obesity and insulin resistance in the offspring, although the mechanisms mediating this effect are unknown. Neither is it known whether maternal leptin affects taste preferences. In this study, we investigated the effect of leptin administration to pregnant mice on the development of diet-induced obesity, food choice, and gene expression in the liver and muscles of the offspring with regard to sex. Leptin was administered to female mice on days 11, 12, and 13 of pregnancy. In male and female offspring, growth rate and intake of standard chow after weaning, obesity development, gene expression in the liver and muscles, and food choice when kept on a high-calorie diet (standard chow, lard, sweet cookies) were recorded. Leptin administration to pregnant females reduced body weight in the female offspring fed on the standard diet. When the offspring were given a high-calorie diet, leptin administration inhibited obesity development and reduced the consumption of cookies only in males. It also increased the consumption of standard chow and the mRNA levels of genes for the insulin receptor and glucose transporter type 4 in the muscles of both male and female offspring. The results demonstrate that an increase in blood leptin levels in pregnant females has a sex-specif ic effect on the metabolism of the offspring increasing resistance to obesity only in male offspring. The mechanism underlying this effect includes a shift in food preference in favor of a balanced diet and maintenance of insulin sensitivity in muscle tissues.

Altered pancreas remodeling following glucose intolerance in pregnancy in mice

Gestational diabetes mellitus increases the risk of dysglycemia postpartum, in part, due to pancreatic β-cell dysfunction. However, no histological evidence exists comparing endocrine pancreas after healthy and glucose-intolerant pregnancies. This study sought to address this knowledge gap, in addition to exploring the contribution of an inflammatory environment to changes in endocrine pancreas after parturition. We used a previously established mouse model of gestational glucose intolerance induced by dietary low protein insult from conception until weaning. Pancreas and adipose samples were collected at 7, 30 and 90 days postpartum for histomorphometric and cytokine analyses, respectively. Glucose tolerance tests were performed prior to euthanasia and blood was collected via cardiac puncture. Pregnant female mice born to dams fed a low protein diet previously shown to develop glucose intolerance at late gestation relative to controls continued to be glucose intolerant until 1 month postpartum. However, glucose tolerance normalized by 3 months postpartum. Glucose intolerance at 7 days postpartum was associated with lower beta- and alpha-cell fractional areas and higher adipose levels of pro-inflammatory cytokine, interleukin-6. By 3 months postpartum, a compensatory increase in the number of small islets and a higher insulin to glucagon ratio likely enabled euglycemia to be attained in the previously glucose-intolerant mice. The results show that impairments in endocrine pancreas compensation in hyperglycemic pregnancy persist after parturition and contribute to prolonged glucose intolerance. These impairments may increase the susceptibility to development of future type 2 diabetes.

Effects of Ocimum sanctum Gestational Administration on Physiological and Behavioral Aspects of Mice Offspring

Aims: Prenatal maternal stress and anxiety affect the offspring causing low birth weight, decreased motor activity, and developmental delays. Ocimum sanctum is cherished as the holiest herb in India and possesses strong adaptogenic, memory enhancer, and anti-stress properties. This study aimed to assess the effect of O. sanctum leaf aqueous extract on preventing pregnancy related stress impact on the mice offspring.&#x0D; Methodology: Pregnant female mice were treated with O. sanctum leaf aqueous extract throughout the gestation period at 100 mg/kg, p.o, while exposed to resident–intruder social stress paradigm. Physiological, morphological, and behavioral aspects of offspring were observed for 6 weeks.&#x0D; Results: Pups of O. sanctum treated mothers showed significantly (P &lt; 0.05) improved body weight, body length, and head length. O. sanctum treatment has significantly reduced eye opening and pinna detachment period (P &lt; 0.05-0.01), and duration for gaining surface righting and mid-air righting refluxes (P &lt; 0.05). O. Sanctum has significantly increased (P &lt; 0.05-0.01) number of correct entries in radial arm maze and line crossing in open field performance.&#x0D; Conclusion: The study outcome proves the beneficial role of O. Sanctum in ameliorating prenatal stress and anxiety induced deleterious effect on offspring.

Preclinical evaluation of immunotherapeutic regimens for fetal/neonatal alloimmune thrombocytopenia

Fetal/neonatal alloimmune thrombocytopenia (FNAIT) is a life-threatening bleeding disorder caused by maternal antibodies directed against paternally inherited antigens present on the surface of fetal platelets. The human platelet alloantigen HPA-1a (formerly known as the PlA1 alloantigen), is the most frequently implicated HPA for causing FNAIT in Whites. A single Leu33Pro amino acid polymorphism residing within the ∼50-amino-acid plexin-semaphorin-integrin domain near the N-terminus of the integrin β3 subunit (platelet membrane glycoprotein IIIa [GPIIIa]) is responsible for generating the HPA-1a and HPA-1b epitopes in human GPIIIa and serves as the central target for alloantibody-mediated platelet destruction. To simulate the etiology of human FNAIT, wild-type female mice were pre-immunized with platelets derived from transgenic mice engineered to express the human HPA-1a epitope on a murine GPIIIa backbone. These mice developed a strong alloimmune response specific for HPA-1a, and when bred with HPA-1a+ males, gave birth to severely thrombocytopenic pups that exhibited an accompanying bleeding phenotype. Administering either polyclonal intravenous immunoglobulin G or a human monoclonal blocking antibody specific for the HPA-1a epitope into pregnant female mice resulted in significant elevation of the neonatal platelet count, normalized hemostasis, and prevented bleeding. The establishment of an alloantigen-specific murine model that recapitulates many of the clinically important features of FNAIT should pave the way for the preclinical development and testing of novel therapeutic and prophylactic modalities to treat or prevent FNAIT in humans.

Identification of the Ghrelin and Cannabinoid CB2 Receptor Heteromer Functionality and Marked Upregulation in Striatal Neurons from Offspring of Mice under a High-Fat Diet.

Cannabinoids have been reported as orexigenic, i.e., as promoting food intake that, among others, is controlled by the so-called “hunger” hormone, ghrelin. The aim of this paper was to look for functional and/or molecular interactions between ghrelin GHSR1a and cannabinoid CB2 receptors at the central nervous system (CNS) level. In a heterologous system we identified CB2-GHSR1a receptor complexes with a particular heteromer print consisting of impairment of CB2 receptor/Gi-mediated signaling. The blockade was due to allosteric interactions within the heteromeric complex as it was reverted by antagonists of the GHSR1a receptor. Cannabinoids acting on the CB2 receptor did not affect cytosolic increases of calcium ions induced by ghrelin acting on the GHSR1a receptor. In situ proximity ligation imaging assays confirmed the expression of CB2-GHSR1a receptor complexes in both heterologous cells and primary striatal neurons. We tested heteromer expression in neurons from offspring of high-fat-diet mouse mothers as they have more risk to be obese. Interestingly, there was a marked upregulation of those complexes in striatal neurons from siblings of pregnant female mice under a high-fat diet.

Effects of Vitamin E on the Epithelial-Mesenchymal Transition in Testicular Development Exposed to Valproic Acid

In testicular differentiation, somatic cells must adopt a specific destiny towards sustentacular, peritubular and interstitial cells, being fundamental for the morphogenesis of seminiferous tubules, mediated by morphogens such as Desert Hedgehog (DHH), insulin-like growth factor-1 (IGF-1) and fibroblastic growth factor 2 (FGF-2). Its alteration could be related to failures in the development mechanisms, such as those caused by valproic acid (VPA), which can be reversed with vitamin E (VE). The objective of the study was to evaluate the epithelial-mesenchymal transition (EMT) in the testicular development of mice exposed to VPA and VE. 12 groups of pregnant female mice were formed that were separated by days post-coital (dpc) at 12.5 dpc, 17.5 dpc and 6 weeks postnatal, each one subdivided into 4 groups of 5 pregnant women each. Subgroups received different treatments from the beginning to the end of gestation orally: 600 mg/kg of VPA, 600 mg/kg of VPA and 200 IU of VE, 200 IU of VE and the control group 0.3 mL of 0.9% physiological solution. Immunohistochemistry was performed for the detection of DHH, IGF-1 and FGF-2. Immunolocalization of DHH was observed in all stages, with more evident significant differences in integrated optical density (IOD) and percentage of immunoreaction area at 6 weeks postnatal, being lower in the VPA group. In IGF-1, lower intensity and distribution of immunostaining was observed in the fetal and pubertal stages in the VPA groups, a similar situation with FGF-2, but only evident at 17.5 dpc, with significant differences. These results demonstrate that VPA can alter EMT between somatic cells in testicular development, with VE being an agent capable of attenuating this process.

Epigenetic Effects Promoted by Neonicotinoid Thiacloprid Exposure.

BackgroundNeonicotinoids, a widely used class of insecticide, have attracted much attention because of their widespread use that has resulted in the decline of the bee population. Accumulating evidence suggests potential animal and human exposure to neonicotinoids, which is a cause of public concern.ObjectivesIn this study, we examined the effects of a neonicotinoid, thiacloprid (thia), on the male reproductive system.MethodsThe pregnant outbred Swiss female mice were exposed to thia at embryonic days E6.5 to E15.5 using “0,” “0.06,” “0.6,” and “6” mg/kg/day doses. Adult male progeny was analyzed for morphological and cytological defects in the testes using hematoxylin and eosin (H&E) staining. We also used immunofluorescence, Western blotting, RT-qPCR and RNA-seq techniques for the analyses of the effects of thia on testis.ResultsWe found that exposure to thia causes a decrease in spermatozoa at doses “0.6” and “6” and leads to telomere defects at all tested doses. At doses “0.6” and “6,” thia exposure leads to an increase in meiotic pachytene cells and a decrease in lumen size, these changes were accompanied by increased testis-to-body weight ratios at high dose. By using RNA-seq approach we found that genes encoding translation, ATP production, ATP-dependent proteins and chromatin-modifying enzymes were deregulated in testes. In addition, we found that exposure to thia results in a decrease in H3K9me3 levels in spermatocytes. The changes in H3K9me3 were associated with a dramatic increase in activity of retroelements.ConclusionOur study suggests that gestational exposure to thia affects epigenetic mechanisms controlling meiosis which could lead to deleterious effects on male spermatogenesis.

Protective role of vitamin E in testicular development of mice exposed to valproic acid.

Valproic acid (VPA) is a teratogenic antiepileptic, causing alterations in oxidative stress in prenatal development, being altered the development of the male reproductive system. The purpose of this study was to determine the protective effect of vitamin E (VE) on the testicular development in embryos, foetuses and pubertal mice exposed to VPA, VPA+VE and only VE. Sixty pregnant adult female mice were used, to which they were administered 600mg/kg of VPA (VPA groups), 600mg/kg of VPA and 200IU of VE (VPA+VE groups), 200IU VE (VE groups) and 0.3ml of 0.9% physiological solution (control groups), showing at 12.5days post-coital (dpc), 17.5dpc and 6weeks postnatal testicular development, and proliferative and apoptotic indices. The groups treated with VPA presented a smaller testicular volume, with greater interstitial space and a delay in the conformation of the testicular cords, shorter lengths and diameters of the germinal epithelium, a smaller number of germline and somatic cells, an increase in cells apoptotic and less proliferation, with significant differences. VE-treated groups behaved similarly to controls. In conclusion, VE reduces the effects caused by VPA throughout testicular development, from embryonic stages, continuing until pubertal stages.